mTOR抑制剂依维莫司在体外对人前列腺癌PC-3细胞株的作用及其机制

发布时间：2018-06-26 04:03

本文选题：依维莫司(Everolimus + RAD001)　；参考：《河北医科大学》2014年硕士论文

【摘要】：目的：在体外研究mTOR抑制剂依维莫司(Everolimus，RAD001)对人前列腺癌PC-3细胞增殖的改变，并探讨其可能的作用机制。方法：体外采用细胞培养技术培养人前列腺癌PC-3细胞株，使用MTT法检测mTOR抑制剂依维莫司(Everolimus，RAD001)1nmol/l干预人前列腺癌PC-3细胞株24h、36h、48h、72h后，PC-3细胞株增殖的变化；流式细胞分析技术（FCM）检测mTOR抑制剂依维莫司(Everolimus，RAD001)干预人前列腺癌PC-3细胞株24h、36h、48h、72h后，细胞周期的变化；Western blot检测mTOR抑制剂依维莫司(Everolimus，RAD001)干预人前列腺癌PC-3细胞株24h、36h、48h、72h后，p-4E-BP1和p-P70-S6K的表达情况。采用SPSS13.0统计软件对实验所得数据分析， P＜0.05有统计学差异。结果： 1MTT结果显示，mTOR抑制剂依维莫司(Everolimus，RAD001)可抑制人前列腺癌PC-3细胞株的增值，且随着药物干预时间的延长，，抑制率逐步增大（P0.01）。 2流式细胞分析技术（FCM）结果显示，人前列腺癌PC-3细胞株经mTOR抑制剂依维莫司(Everolimus，RAD001)干预后，随着药物干预时间的延长，G0/G1期的细胞百分比逐步升高，S、G2/M期的细胞百分比逐步降低，使绝大部分PC-3细胞被中断在细胞周期的G0/G1期（P＜0.01）。 3Western blot检测结果显示，mTOR抑制剂依维莫司(Everolimus，RAD001)干预人前列腺癌PC-3细胞株后，随着药物干预时间的延长，p-4E-BP1和p-P70-S6K的表达逐步被抑制（P0.01）。结论： mTOR抑制剂依维莫司(Everolimus，RAD001)能够抑制人前列腺癌PC-3细胞株的增值，其机制可能与PI3K/Akt/mTOR信号传导通路中的mTOR被抑制，继而抑制了其下游的两个重要蛋白（即翻译抑制因子eIF-4E结合蛋白1(4E-BP1)和核糖体蛋白P70-S6K）的磷酸化有关。
[Abstract]:Aim: to investigate the effect of Everolimus RAD001, an inhibitor of mTOR, on the proliferation of human prostate cancer cell line PC-3 in vitro and to explore its possible mechanism. Methods: human prostate cancer PC-3 cell line was cultured by cell culture technique in vitro. The proliferation of human prostate cancer PC-3 cell line was measured by MTT assay after treatment with 1nmol/l for 24 h, 36 h and 48 h, respectively. Flow cytometry (FCM) was used to detect the expression of p-4E-BP1 and p-P70-S6K in human prostate cancer PC-3 cells treated with Everolimus RAD001, a mTOR inhibitor, for 24 h or 36 h or 48 h or 72 h. The changes of cell cycle were detected by Western blot. The expression of p-4E-BP1 and p-P70-S6K in human prostate cancer cell line PC-3 treated with Everolimusine RAD001 for 24 h, 36 h, 48 h and 72 h later. SPSS 13.0 statistical software was used to analyze the experimental data (P < 0.05). Results: 1MTT results showed that Everolimus RAD001, an inhibitor of mTOR, inhibited the proliferation of human prostate cancer cell line PC-3. The inhibition rate increased gradually (P0.01). 2 the results of flow cytometry (FCM) showed that human prostate cancer PC-3 cell line was treated with the mTOR inhibitor Everolimus ARAD001. The percentage of cells in G _ 0 / G _ 1 phase increased gradually with the prolongation of drug intervention time, and the percentage of cells in G _ 2 / M phase decreased gradually. The majority of PC-3 cells were interrupted in G0 / G1 phase of cell cycle (P < 0. 01). 3 the results of Western blot analysis showed that human prostate cancer PC-3 cell line was treated with Everolimus Rad001, an inhibitor of mTOR. The expression of p-4E-BP1 and p-P70-S6K was inhibited gradually with the prolongation of drug intervention time (P0.01). Conclusion: Everolimus RAD001, an inhibitor of mTOR, can inhibit the proliferation of human prostate cancer cell line PC-3, and its mechanism may be related to the inhibition of mTOR in the PI3K / Akt / mTOR signaling pathway. The phosphorylation of two important proteins downstream, the translation inhibitory factor eIF-4E binding protein 1 (4E-BP1) and ribosomal protein P70-S6K, was subsequently inhibited.
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.25

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